I want to make the standard curve for my acetic acid adsorption data on HPLC. I already measured the acetic acid from my reactors using the HPLC but haven't measured the right standard curve. I tried using some references from the web but was not pretty much confident of it. Does anyone know how to make a reliable standard curve for acetic acid on an HPLC device (I am using a reagent of H2SO4 5 mM)?
8/17/23
Dear Dick Dick,
In our lab, we frequently measure the components of wine, which contains small amounts of acetic acid (HAc). We do this by running a series (5 or 6) of HAc standards on our HPLC, along with our unknown samples. I can't recall the column we use, but it may be similar or identical to yours, as we use 5 mM H2SO4 as the mobile phase. After the run, we call up chromatographic profiles of the standards and integrate the HAc peak areas, then plot this vs. the HAc concentrations of the standards to get a std curve. R2 is usually ~0.98 or higher. We also integrate the HAc peak areas of the unknowns. After that, it is a matter of getting the HPLC's computer to calculate the concentration of HAc in the unknowns.
We use an Agilent HPLC. Usually, the device automatically integrates the peaks, but occasionally, depending on the baseline, we have to go back and manually integrate them. The computer software will do the calculation for you after the std curve is plotted. If your HPLC can't do this, you can also prepare a standard curve in Excel. Both ways work.
I hope this information helps you.
Bill Colonna Iowa State University, Ames, Iowa, USA [email protected]
Hello Problems with calibrations? You just need to decrease the uncertainty of your Analytical method. For example, use larger volume flasks and pipettes (lower degree of error). Also the plot of residuals shows which level of your curve has more error.
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